{"artifact":{"id":"experiment-exp-debate-bb0a058a7032","artifact_type":"experiment","entity_ids":null,"title":"Proposed experiment from debate on Perivascular spaces and glymphatic clearance failure in AD","quality_score":0.675,"created_by":"crosslink-v2","provenance_chain":null,"content_hash":null,"metadata":{"source":{"type":"manual","source_name":"debate_extraction","extracted_by":"backfill_v1","extraction_date":"2026-04-16T01:00:16.909563Z"},"summary":"# Proposed experiment from debate on Perivascular spaces and glymphatic clearance failure in AD\n## Background and Rationale\nThis experiment investigates the mechanistic relationship between TREK-1 potassium channels and AQP4 polarization in the context of glymphatic clearance dysfunction associated with Alzheimer's disease. TREK-1 channels are mechanosensitive potassium channels expressed in astrocytes that respond to mechanical stimuli and may regulate astrocytic volume and function. The scient","entities":{"genes":["TREK"],"diseases":["Neurodegeneration"]},"model_system":"mouse","_schema_version":1,"experiment_type":"falsification","primary_outcome":"Real-time measurement of AQP4-GFP polarization changes at astrocytic endfeet following selective TREK-1 channel activation, correlated with patch-clamp recordings of membrane conductance.","methodology_notes":"**Phase 1: Animal Preparation and Baseline Characterization (Days 1-7)**\n• Obtain 8-10 week old AQP4-GFP transgenic mice (n=48, equal male/female distribution)\n• Perform baseline cognitive assessment using Morris water maze and novel object recognition\n• Collect baseline CSF samples via cisterna magna puncture for Aβ40/42 and tau measurements\n• Establish baseline glymphatic function using fluorescent tracer injection (Texas Red 3kDa dextran)\n• Randomize animals into treatment groups: vehicle control (n=12), TREK-1 agonist ML335 (10mg/kg, n=12), TREK-1 antagonist spadin (5mg/kg, n=12), AD model + ML335 (n=12)\n\n**Phase 2: Real-time AQP4 Polarization Imaging (Days 8-14)**\n• Prepare acute brain slices (300μm thickness) from cortex and hippocampus\n• Mount slices in perfusion chamber with oxygenated aCSF at 32°C\n• Apply TREK-1 modulators via perfusion system (ML335 100nM, spadin 1μM)\n• Perform two-photon microscopy imaging of AQP4-GFP at perivascular endfeet\n• Quantify AQP4 polarization inde","replication_status":"single_study","extraction_metadata":{"backfill_at":"2026-04-16T01:00:16.909569","needs_review":true,"extraction_notes":"Backfilled from debate_extraction source (no PMID 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